• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.506-514
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• 2020

4-Hydroxybenzyl alcohol (4-HB alcohol) is one of the major active components of Gastrodia elata Blume, with beneficial effects on neurological disorders such as headache, convulsive behavior, and dizziness. Here, we developed a metabolically engineered Corynebacterium glutamicum strain able to produce 4-HB alcohol from 4-hydroxybenzoate (4-HBA). First, the strain APS963 was obtained from the APS809 strain via the insertion of aroK from Methanocaldococcus jannaschii into the NCgl2922-deleted locus. As carboxylic acid reductase from Nocardia iowensis catalyzes the reduction of 4HBA to 4-hydroxybenzaldehyde (4-HB aldehyde), we then introduced a codon-optimized car gene into the genome of APS963, generating the GAS177 strain. Then, we deleted creG coding for a putative short-chain dehydrogenase and inserted ubiCpr encoding a product-resistant chorismate-pyruvate lyase into the pcaHG-deleted locus. The resulting engineered GAS355 strain accumulated 2.3 g/l 4-HB alcohol with 0.32 g/l 4-HBA and 0.3 g/l 4-HB aldehyde as byproducts from 8% glucose after 48 h of culture.

• Animal Bioscience
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• v.35 no.2
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• pp.204-216
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• 2022

Objective: The aim of this study was to investigate the effects of dietary supplementation with a multi-strain probiotic (MSP) product containing of Bifidobacterium animalis, Lactobacillus casei, Streptococcus faecalis, and Bacillus cerevisiae on growth, health, and fecal bacterial composition of dairy calves during the first month of life. Methods: Forty Holstein calves (24 female and 16 male) at 2 d of age were grouped by sex and date of birth then randomly assigned to 1 of 4 treatments: milk replacer supplementation with 0 g (0MSP), 2 g (2MSP), 4 g (4MSP), and 6 g (6MSP) MSP per calf per day. Results: Supplementation of MSP did not result in any significant differences in parameters of body measurements of calves during the 30 d period. As the dosage of MSP increased, the average daily gain (p = 0.025) and total dry matter intake (p = 0.020) of calves showed a linear increase. The fecal consistency index of the 2MSP, 4MSP, and 6MSP group calves were lower than that of the 0MSP group calves (p = 0.003). As the dosage of MSP increased, the concentrations of lactate dehydrogenase (p = 0.068) and aspartate aminotransferase (p = 0.081) in serum tended to decrease, whereas the concentration of total cholesterol increased quadratically (p = 0.021). The relative abundance of Dorea in feces was lower (p = 0.011) in the 2MSP, 4MSP, and 6MSP group calves than that in the 0MSP group calves. The relative abundance of Dorea (p = 0.001), Faecalibacterium (p = 0.050), and Mitsuokella (p = 0.030) decreased linearly, whereas the relative abundance of Prevotella tended to increase linearly as the dosage of MSP increased (p = 0.058). Conclusion: The MSP product can be used to reduce the diarrhea, improve the performance, and alter the composition of the fecal bacteria in neonatal dairy calves under the commercial conditions.

• Animal Bioscience
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• v.35 no.11
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• pp.1787-1799
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• 2022

Objective: Choline deficiency, one main trigger for nonalcoholic fatty liver disease (NAFLD), is closely related to lipid metabolism disorder. Previous study in a choline-deficient model has largely focused on gene expression rather than gene structure, especially sparse are studies regarding to alternative splicing (AS). In modern life science research, primary hepatocytes culture technology facilitates such studies, which can accurately imitate liver activity in vitro and show unique superiority. Whereas limitations to traditional hepatocytes culture technology exist in terms of efficiency and operability. This study pursued an optimization culture method for duck primary hepatocytes to explore AS in choline-deficient model. Methods: We performed an optimization culture method for duck primary hepatocytes with multi-step digestion procedure from Pekin duck embryos. Subsequently a NAFLD model was constructed with choline-free medium. RNA-seq and further analysis by rMATS were performed to identify AS events alterations in choline-deficency duck primary hepatocytes. Results: The results showed E13 (embryonic day 13) to E15 is suitable to obtain hepatocytes, and the viability reached over 95% by trypan blue exclusion assay. Primary hepatocyte retained their biological function as well identified by Periodic Acid-Schiff staining method and Glucose-6-phosphate dehydrogenase activity assay, respectively. Meanwhile, genes of alb and afp and specific protein of albumin were detected to verify cultured hepatocytes. Immunofluorescence was used to evaluate purity of hepatocytes, presenting up to 90%. On this base, choline-deficient model was constructed and displayed significantly increase of intracellular triglyceride and cholesterol as reported previously. Intriguingly, our data suggested that AS events in choline-deficient model were implicated in pivotal biological processes as an aberrant transcriptional regulator, of which 16 genes were involved in lipid metabolism and highly enriched in glycerophospholipid metabolism. Conclusion: An effective and rapid protocol for obtaining duck primary hepatocytes was established, by which our findings manifested choline deficiency could induce the accumulation of lipid and result in aberrant AS events in hepatocytes, providing a novel insight into various AS in the metabolism role of choline.

• Nutrition Research and Practice
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• v.16 no.1
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• pp.33-45
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• 2022

BACKGROUND/OBJECTIVES: Ginseng extract (GSE) and taurine (TR) are widely used antifatigue resources in functional foods. However, the mechanism underlying the antifatigue effects of GSE and TR are still unclear. Hence, we investigated whether GSE and TR have synergistic effects against fatigue in mice. MATERIALS/METHODS: L6 cells were treated with different concentrations of TR and GSE, and cell viability was determined using 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium. Oxidative stress was analyzed by immunocytochemistry using MitoTracker^TM Red FM and an anti-8-oxoguanine antibody. Respiratory gas analysis was performed to investigate metabolism. Expression of an activated protein kinase was analyzed using immunohistochemistry. Gene expression of cluster of differentiation 36 and pyruvate dehydrogenase lipoamide kinase isozyme 4 was measured using reverse transcription-polymerase chain reaction. Mice were orally administered TR, GSE, or their combination for 30 days, and then fatigue-related parameters, including lactate, blood urea nitrogen, and glycogen, were measured after forced swimming. RESULTS: TR and GSE reduced oxidative stress levels in hydrogen peroxide-stimulated L6 cells and enhanced the oxygen uptake and lipid metabolism in mice after acute exercise. After oral administration of TR or GSE for 30 days, the fatigue-related parameters did not change in mice. However, the mice administered GSE (400 mg/kg/day) alone for 30 days could swim longer than those from the other groups. Further, no synergistic effect was observed after the swimming exercise in mice treated with the TR and GSE combination for 30 days. CONCLUSIONS: Taken together, our data suggest that TR and GSE may exert antifatigue effects in mice after acute exercise by enhancing oxygen uptake and lipid oxidation.

• Korean Journal of Clinical Laboratory Science
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• v.54 no.2
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• pp.103-109
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• 2022

In December 2019, the coronavirus disease 2019 (COVID-19) caused by the virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in China and spread rapidly around the world, infecting millions of people. Cases of COVID-19 infection were observed to lead to viral pneumonia. Thirty-five patients admitted to the Gyeonggi Medical Center, South Korea, between November 2020 to January 2021, were found to have been infected with the influenza virus A and B, which cause symptoms similar to COVID-19. The records of these patients and those of COVID-19 patients who visited the hospital for medical examination were compared. The study patients included thirty patients with COVID-19 and/or influenza, five of those with influenza alone. A group of 121 patients without infection was used as control. Patients with COVID-19 and influenza had significantly higher lactate dehydrogenase levels than the patients with COVID-19 alone. The erythrocyte sedimentation rate (ESR) was higher in patients with COVID-19 alone than in other groups. Significant clinical outliers were observed in the COVID-19 and influenza infection group compared with the COVID-19 alone group. These results are expected to play an important role in the analysis of the hematological data of infected patients and the comparison of simultaneous and single infection data to determine clinical symptoms and other signs. These results may also assist in the development of vaccines and treatments for COVID-19.

• The Korea Journal of Herbology
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• v.36 no.6
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• pp.1-8
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• 2021

Objectives : Atractylodis rhizoma Alba has been traditionally used as a medicinal resource that is used for enhancing Qi (氣) in traditional medicine in Korea, China, and Japan. This study investigated the protective effects of Atractylodis rhizoma Alba extract (ARE) against trimethyltin (TMT), a neurotoxin that causes selective hippocampal injury, using both in vitro and in vivo models. Methods : We investigated the effects of ARE on TMT- (5mM) induced cytotoxicity in primary cultures of mouse hippocampal cells (7 days in vitro ) and on hippocampal injury in C57BL/6 mice injected with TMT (2.6 mg/kg). Results : We observed that ARE treatment (0 - 50 ㎍/mL) significantly reduced TMT-induced cytotoxicity in cultured hippocampal neurons in a dose-dependent manner, based on results of lactate dehydrogenase and 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide assays. Additionally, this study showed that orally administered ARE (5 mg/kg; between -6 and 0 days before TMT injection) significantly attenuated seizures in adult mice. Furthermore, quantitative analysis of allograft inflammatory factor-1 (Iba-1)- and glial fibrillary acidic protein (GFAP)- positive cells showed significantly reduced levels of Iba-1- and GFAP-positive cell bodies in the dentate gyrus of mice treated with ARE prior to TMT injection. These findings indicate the significant protective effects of ARE against the TMT-induced massive activation of microglia and astrocytes in the hippocampus. Conclusions : We conclude that ARE minimizes the detrimental effects of TMT-induced hippocampal neurotoxicity, both in vitro and in vivo . Our findings may serve as useful guidelines to support ARE administration as a promising pharmacotherapeutic approach to hippocampal degeneration.

• Herbal Formula Science
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• v.30 no.2
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• pp.85-93
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• 2022

Objectives : This study investigated anti-inflammatory effects of Nardotidis seu Sulculii Concha water extract (NSCE) against restraint-induced stress. Methods : In vivo, NSCE was orally administered to male white mice at concentrations of 250 mg/kg and 500 mg/kg for 3 days, and then restraint-induced stress was induced for 6 hours. The level of liver damage was measured by serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH). The stress-related hormones such as cortisol and corticosterone were measured by ELISA assay. Also, western blot analysis was performed to detect expression of mitogen-activated protein kinase (MAPK) and cyclooxygenase-2 (COX-2) proteins. Pathological changes were observed by hematoxylin and eosin (H&E) staining of the liver tissue, and Immunohistochemical (IHC) staining was performed to examine liver inflammation through macrophage infiltration. Results : The AST, ALT, LDH and the stress related hormones such as cortisol and corticosterone were significantly decreased in the NSCE treated group compared with stress group. In histological analysis, H&E staining of liver tissues did not detect the hepatic injury or damage in all groups. As a result of IHC staining, it was confirmed that infiltration of macrophages was increased in the stress-induced group, but decreased in the group treated with NSCE. The COX-2 and MAPK proteins expression was significantly increased by restraint-induced stress, but these proteins were decreased in the NSCE treated group. Conclusions : These results suggest that NSCE has the anti-inflammatory activity in restraint-induced stress model, and it is believed that NSCE can be used for the prevention of liver inflammation.

• Animal Bioscience
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• v.35 no.5
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• pp.763-777
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• 2022

Objective: Excessive lipid accumulation in adipocytes results in prevalence of obesity and metabolic syndrome. Curcumin (CUR), a naturally phenolic active ingredient, has been shown to have lipid-lowering effects. However, its underlying mechanisms have remained largely unknown. Therefore, the study aims to determine the effect of CUR on cellular lipid accumulation in porcine subcutaneous preadipocytes (PSPA) and to clarify novel mechanisms. Methods: The PSPA were cultured and treated with or without CUR. Both cell counting Kit-8 and lactate dehydrogenase release assays were used to examine cytotoxicity. Intracellular lipid contents were measured by oil-red-o staining extraction and triglyceride quantification. Apoptosis was determined by flow cytometry and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling assay. Adipogenic and apoptosis genes were analyzed by quantitative polymerase chain reaction and Western blot. Results: The CUR dose-dependently reduced the proliferation and lipid accumulation of PSPA. Noncytotoxic doses of CUR (10 to 20 μM) significantly inhibited extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation and expression of adipogenic genes peroxisome proliferation-activity receptor-γ (PPAR-γ), CCAAT/enhancer binding protein-α, sterol regulatory element-binding protein-1c, adipocyte protein-2, glucose transporter-4 as well as key lipogenic enzymes fatty acid synthase and acetyl-CoA carboxylase, while ERK1/2 activation significantly reversed CUR-reduced lipid accumulation by increasing PPAR-γ. Furthermore, compared with differentiation induced media treated cells, higher dose of CUR (30 μM) significantly decreased the expression of AKT and B-cell lymphoma-2 (BCL-2), while increased the expression of BCL-2-associated X (BAX) and the BAX/BCL-2 expression ratio, suggesting triggered apoptosis by inactivating AKT and increasing BAX/BCL-2 ratio and Caspase-3 expression. Moreover, AKT activation significantly rescued CUR inhibiting lipid accumulation via repressing apoptosis. Conclusion: These results demonstrate that CUR is capable of suppressing differentiation by inhibiting ERK1/2-PPAR-γ signaling pathway and triggering apoptosis via decreasing AKT and subsequently increasing BAX/BCL-2 ratio and Caspase-3, suggesting that CUR provides an important method for the reduction of porcine body fat, as well as the prevention and treatment of human obesity.

• The Korea Journal of Herbology
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• v.34 no.6
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• pp.45-55
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• 2019

Objective : Diabetic gastroparesis is a complication that is defined as delayed gastric emptying and upper gastrointestinal symptoms and often occurs in long-standing diabetic patients. Bojungikgi-tang (BJT) is a traditional oriental herbal formula that has long been used for the treatment of digestive disorders. The purpose of this study was to investigate the effects of BJT on streptozotocin (STZ)-induced diabetic gastroparesis rat model. Methods : Sprague-Dawley (SD) male rats (250-270g) were divided into 13 groups including normal group, STZ-induced diabetic control group, BJT diet (7 various concentrations), and insulin-, glibenclamide-, metformin-treated group were used for the experiments for the comparison. Diabetic gastroparesis was induced by intraperitoneal injection of STZ. The water intake, food intake, body weights and fasting blood glucose levels were measured. After 4 weeks the animals were sacrificed and gastrin, leptin, insulin, hemoglobin A1C (HbA1c), lactate, lactate dehydrogenase (LDH), bilirubin, creatinine, albumin and lipid levels were evaluated. Results : Intraperitoneal injection of BJT for 4 weeks resulted in increased levels of gastrin in blood and decreased leptin and lactate concentration in STZ-induced diabetic gastroparesis rat model. BJT did not affect insulin, fasting glucose, HbA1c, and lipid levels in STZ-induced diabetic gastroparesis rat model. Conclusion : These results indicated that BJT would have protect effect on diabetic gastroparesis through the improvement effect of gastric motility and fatigue syndrome in STZ-induced diabetic rats. This study shows that BJT might be effective for treatment of diabetes and its complications such as gastroparesis.

• The Korea Journal of Herbology
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• v.35 no.1
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• pp.45-55
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• 2020

Objectives : The purpose of this study is to determine whether Rhus verniciflua Stokes with Latin name Lacca Sinica Exsiccata, and Eucommia ulmoides Oliver with Latin name Eucommiae Cortex Extract Combination (ILF-RE) improves laboratory test results in participants with liver function disorder. Methods : This study was conducted at Woosuk university Korean medicine hospital where participants with high serum alanine transaminase (ALT) levels from 45 to 135 U/L were enrolled. Subjects received ILF-RE 3.6 g (1.2 g/day as ILF-RE) or placebo 3.6 g for 12 weeks. It was confirmed that urushiol was not detected in ILF-RE. The primary outcomes were the decrement degree of serum ALT and gamma-glutamyl transferase (GGT) levels between two groups. The secondary outcomes were the decrement degree of serum aspartate transaminase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LD), total bilirubin, total cholesterol, triglyceride (TG) and fatty liver index (FLI) levels between two groups. Adverse events, skin prick tests, laboratory tests, and vital signs were observed and analyzed to confirm the safety of ILF-RE.1) Results : In the ILF-RE group, the liver function index ALT, GGT, lipid metabolism index TG, and fatty liver index FLI were significantly decreased compared to the placebo group. There was no significant difference in ILF-RE group in terms of adverse events, severe adverse events, skin prick test, laboratory test, and vital signs compared with placebo group. Conclusions : ILF-RE was found to be effective in improving liver function. In addition, no clinically significant adverse events or body changes were observed during this study.